摘要
利用Caco-2细胞和HaCaT细胞建立肠-皮肤共培养体系,采用免疫荧光法评价安石榴苷对中波紫外线(UVB)诱导的光损伤产物环丁烷嘧啶二聚体(CPD)的抑制效果,通过实时荧光定量PCR、免疫沉淀、蛋白免疫印迹法探究安石榴苷对核苷酸切除修复(NER)途径相关基因转录和产物表达的影响。结果表明,安石榴苷能降低HaCaT细胞中的CPD水平,上调着色性干皮病基因组C(XPC)的mRNA转录水平,并以SIRT1基因依赖的方式降低XPA蛋白的乙酰化水平。这表明在Caco-2细胞与HaCaT细胞的肠-皮肤共培养体系中,安石榴苷可能通过激活NER途径相关的XPC/XPA蛋白促进UVB诱导的光损伤的修复,其作用依赖于SIRT1蛋白的去乙酰化活性。
In this study, the co-culture system of Caco-2 and HaCaT cells was established, and the inhibitory effect of punicalagin on cyclobutene pyrimidine dimers(CPD) which was one major type of UVB-induced DNA lesions was evaluated by immunofluorescence. Then the effects of punicalagin on transcription and expression levels of nucleotide excision repair(NER) related genes was further investigated by quantitative real-time PCR, immunoprecipitation and western-blotting. The results showed that punicalagin reduced CPD level in HaCaT cells, upregulated xeroderma pigmentosum group C(XPC) mRNA level and attenuated XPA acetylation in a SIRT1-dependent manner. This showed, punicalagin′s efficacy against UVB-induced photodamage in the co-culture system of Caco-2 and HaCaT cells was probably activated by the NER related XPC/XPA which depended on the deacetylation activity of SIRT1.
作者
沈芳铭
尹淑涛
赵冲
SHEN Fangming;YIN Shutao;ZHAO Chong(College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083,China)
出处
《轻工学报》
北大核心
2022年第6期50-57,共8页
Journal of Light Industry
基金
国家自然科学基金青年科学基金项目(31901691)
